Friedman Lisa, Kolter Roberto
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115, USA.
Mol Microbiol. 2004 Feb;51(3):675-90. doi: 10.1046/j.1365-2958.2003.03877.x.
Pseudomonas aeruginosa forms diverse matrix-enclosed surface-associated multicellular assemblages (biofilms) that aid in its survival in a variety of environments. One such biofilm is the pellicle that forms at the air-liquid interface in standing cultures. We screened for transposon insertion mutants of P. aeruginosa PA14 that were unable to form pellicles. Analysis of these mutants led to the identification of seven adjacent genes, named pel genes, the products of which appear to be involved in the formation of the pellicle's extracellular matrix. In addition to being required for pellicle formation, the pel genes are also required for the formation of solid surface-associated biofilms. Sequence analyses predicted that three pel genes encode transmembrane proteins and that five pel genes have functional homologues involved in carbohydrate processing. Microscopic and macroscopic observations revealed that wild-type P. aeruginosa PA14 produces a cellulase-sensitive extracellular matrix able to bind Congo red; no extracellular matrix was produced by the pel mutants. A comparison of the carbohydrates produced by the wild-type strain and pel mutants suggested that glucose was a principal component of the matrix material. Together, these results suggest that the pel genes are responsible for the production of a glucose-rich matrix material required for the formation of biofilms by P. aeruginosa PA14.
铜绿假单胞菌形成多种被基质包裹的、与表面相关的多细胞聚集体(生物膜),这有助于其在各种环境中生存。其中一种生物膜是在静置培养物的气液界面形成的菌膜。我们筛选了不能形成菌膜的铜绿假单胞菌PA14转座子插入突变体。对这些突变体的分析导致鉴定出七个相邻基因,命名为pel基因,其产物似乎参与菌膜细胞外基质的形成。除了菌膜形成所需外,pel基因也是固体表面相关生物膜形成所必需的。序列分析预测,三个pel基因编码跨膜蛋白,五个pel基因具有参与碳水化合物加工的功能同源物。显微镜和宏观观察表明,野生型铜绿假单胞菌PA14产生一种能结合刚果红的对纤维素酶敏感的细胞外基质;pel突变体不产生细胞外基质。对野生型菌株和pel突变体产生的碳水化合物的比较表明,葡萄糖是基质材料的主要成分。这些结果共同表明,pel基因负责产生铜绿假单胞菌PA14形成生物膜所需的富含葡萄糖的基质材料。